105 results on '"Robbie G. Majzner"'
Search Results
2. CAR T Cell Therapy for Neuroblastoma
- Author
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Rebecca M. Richards, Elena Sotillo, and Robbie G. Majzner
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neuroblastoma ,pediatric oncology ,immunotherapy ,CAR T cells ,adoptive T cell therapy ,clinical trials ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Patients with high risk neuroblastoma have a poor prognosis and survivors are often left with debilitating long term sequelae from treatment. Even after integration of anti-GD2 monoclonal antibody therapy into standard, upftont protocols, 5-year overall survival rates are only about 50%. The success of anti-GD2 therapy has proven that immunotherapy can be effective in neuroblastoma. Adoptive transfer of chimeric antigen receptor (CAR) T cells has the potential to build on this success. In early phase clinical trials, CAR T cell therapy for neuroblastoma has proven safe and feasible, but significant barriers to efficacy remain. These include lack of T cell persistence and potency, difficulty in target identification, and an immunosuppressive tumor microenvironment. With recent advances in CAR T cell engineering, many of these issues are being addressed in the laboratory. In this review, we summarize the clinical trials that have been completed or are underway for CAR T cell therapy in neuroblastoma, discuss the conclusions and open questions derived from these trials, and consider potential strategies to improve CAR T cell therapy for patients with neuroblastoma.
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- 2018
- Full Text
- View/download PDF
3. 244 Targeting gangliosides in pediatric cancer
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Min Huang, Guillermo N Dalton, Won-Ju Kim, Nathaniel W Mabe, Maria Caterina Rotiroti, Aidan M Tousley, Kimberly Stegmaier, and Robbie G Majzner
- Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Published
- 2023
- Full Text
- View/download PDF
4. Tumor inflammation-associated neurotoxicity
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Jasia Mahdi, Jorg Dietrich, Karin Straathof, Claire Roddie, Brian J. Scott, Tom Belle Davidson, Laura M. Prolo, Tracy T. Batchelor, Cynthia J. Campen, Kara L. Davis, Juliane Gust, Michael Lim, Robbie G. Majzner, Julie R. Park, Sonia Partap, Sneha Ramakrishna, Rebecca Richards, Liora Schultz, Nicholas A. Vitanza, Leo D. Wang, Crystal L. Mackall, and Michelle Monje
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General Medicine ,General Biochemistry, Genetics and Molecular Biology - Published
- 2023
5. Co-opting signalling molecules enables logic-gated control of CAR T cells
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Aidan M. Tousley, Maria Caterina Rotiroti, Louai Labanieh, Lea Wenting Rysavy, Won-Ju Kim, Caleb Lareau, Elena Sotillo, Evan W. Weber, Skyler P. Rietberg, Guillermo Nicolas Dalton, Yajie Yin, Dorota Klysz, Peng Xu, Eva L. de la Serna, Alexander R. Dunn, Ansuman T. Satpathy, Crystal L. Mackall, and Robbie G. Majzner
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Multidisciplinary - Published
- 2023
6. Reprogramming Cancer into Antigen-Presenting Cells as a Novel Immunotherapy
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Miles H. Linde, Amy C. Fan, Thomas Köhnke, Aaron C. Trotman-Grant, Sarah F. Gurev, Paul Phan, Feifei Zhao, Naomi L. Haddock, Kevin A. Nuno, Eric J. Gars, Melissa Stafford, Payton L. Marshall, Christopher G. Dove, Ian L. Linde, Niklas Landberg, Lindsay P. Miller, Robbie G. Majzner, Tian Yi Zhang, and Ravindra Majeti
- Subjects
Oncology - Abstract
Therapeutic cancer vaccination seeks to elicit activation of tumor-reactive T cells capable of recognizing tumor-associated antigens (TAA) and eradicating malignant cells. Here, we present a cancer vaccination approach utilizing myeloid-lineage reprogramming to directly convert cancer cells into tumor-reprogrammed antigen-presenting cells (TR-APC). Using syngeneic murine leukemia models, we demonstrate that TR-APCs acquire both myeloid phenotype and function, process and present endogenous TAAs, and potently stimulate TAA-specific CD4+ and CD8+ T cells. In vivo TR-APC induction elicits clonal expansion of cancer-specific T cells, establishes cancer-specific immune memory, and ultimately promotes leukemia eradication. We further show that both hematologic cancers and solid tumors, including sarcomas and carcinomas, are amenable to myeloid-lineage reprogramming into TR-APCs. Finally, we demonstrate the clinical applicability of this approach by generating TR-APCs from primary clinical specimens and stimulating autologous patient-derived T cells. Thus, TR-APCs represent a cancer vaccination therapeutic strategy with broad implications for clinical immuno-oncology. Significance: Despite recent advances, the clinical benefit provided by cancer vaccination remains limited. We present a cancer vaccination approach leveraging myeloid-lineage reprogramming of cancer cells into APCs, which subsequently activate anticancer immunity through presentation of self-derived cancer antigens. Both hematologic and solid malignancies derive significant therapeutic benefit from reprogramming-based immunotherapy. This article is highlighted in the In This Issue feature, p. 1027
- Published
- 2023
7. Time to resolution of iodine‐123 metaiodobenzylguanidine ( <scp> 123 I‐MIBG </scp> ) avidity and local control outcomes for high‐risk neuroblastoma following radiation therapy
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Justin Oh, Paulina Gutkin, Yi Peng Wang, Navjot Sandhu, Robbie G Majzner, Helen Nadel, Hiroyuki Shimada, Olivia Lansinger, Rie von Eyben, Sarah Donaldson, Matias Bruzoni, Quaovi H Sodji, and Susan M Hiniker
- Subjects
Oncology ,Radiology, Nuclear Medicine and imaging - Published
- 2022
8. Transition to a mesenchymal state in neuroblastoma confers resistance to anti-GD2 antibody via reduced expression of ST8SIA1
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Nathaniel W. Mabe, Min Huang, Guillermo N. Dalton, Gabriela Alexe, Daniel A. Schaefer, Anna C. Geraghty, Amanda L. Robichaud, Amy S. Conway, Delan Khalid, Marius M. Mader, Julia A. Belk, Kenneth N. Ross, Michal Sheffer, Miles H. Linde, Nghi Ly, Winnie Yao, Maria Caterina Rotiroti, Benjamin A. H. Smith, Marius Wernig, Carolyn R. Bertozzi, Michelle Monje, Constantine S. Mitsiades, Ravindra Majeti, Ansuman T. Satpathy, Kimberly Stegmaier, and Robbie G. Majzner
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Neuroblastoma ,Cancer Research ,Oncology ,Gangliosides ,Antibodies, Monoclonal ,Humans ,Immunotherapy ,Neoplasm Recurrence, Local ,Child - Abstract
Immunotherapy with anti-GD2 antibodies has advanced the treatment of children with high-risk neuroblastoma, but nearly half of patients relapse, and little is known about mechanisms of resistance to anti-GD2 therapy. Here, we show that reduced GD2 expression was significantly correlated with the mesenchymal cell state in neuroblastoma and that a forced adrenergic-to-mesenchymal transition (AMT) conferred downregulation of GD2 and resistance to anti-GD2 antibody. Mechanistically, low-GD2-expressing cell lines demonstrated significantly reduced expression of the ganglioside synthesis enzyme ST8SIA1 (GD3 synthase), resulting in a bottlenecking of GD2 synthesis. Pharmacologic inhibition of EZH2 resulted in epigenetic rewiring of mesenchymal neuroblastoma cells and re-expression of ST8SIA1, restoring surface expression of GD2 and sensitivity to anti-GD2 antibody. These data identify developmental lineage as a key determinant of sensitivity to anti-GD2 based immunotherapies and credential EZH2 inhibitors for clinical testing in combination with anti-GD2 antibody to enhance outcomes for children with neuroblastoma.
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- 2022
9. Supplemental Figure S9 from Reprogramming Cancer into Antigen-Presenting Cells as a Novel Immunotherapy
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Ravindra Majeti, Tian Yi Zhang, Robbie G. Majzner, Lindsay P. Miller, Niklas Landberg, Ian L. Linde, Christopher G. Dove, Payton L. Marshall, Melissa Stafford, Eric J. Gars, Kevin A. Nuno, Naomi L. Haddock, Feifei Zhao, Paul Phan, Sarah F. Gurev, Aaron C. Trotman-Grant, Thomas Köhnke, Amy C. Fan, and Miles H. Linde
- Abstract
Primary B-ALL TR-APCs can be generated via exogenous cytokine stimulation
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- 2023
10. Data from Reprogramming Cancer into Antigen-Presenting Cells as a Novel Immunotherapy
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Ravindra Majeti, Tian Yi Zhang, Robbie G. Majzner, Lindsay P. Miller, Niklas Landberg, Ian L. Linde, Christopher G. Dove, Payton L. Marshall, Melissa Stafford, Eric J. Gars, Kevin A. Nuno, Naomi L. Haddock, Feifei Zhao, Paul Phan, Sarah F. Gurev, Aaron C. Trotman-Grant, Thomas Köhnke, Amy C. Fan, and Miles H. Linde
- Abstract
Therapeutic cancer vaccination seeks to elicit activation of tumor-reactive T cells capable of recognizing tumor-associated antigens (TAA) and eradicating malignant cells. Here, we present a cancer vaccination approach utilizing myeloid-lineage reprogramming to directly convert cancer cells into tumor-reprogrammed antigen-presenting cells (TR-APC). Using syngeneic murine leukemia models, we demonstrate that TR-APCs acquire both myeloid phenotype and function, process and present endogenous TAAs, and potently stimulate TAA-specific CD4+ and CD8+ T cells. In vivo TR-APC induction elicits clonal expansion of cancer-specific T cells, establishes cancer-specific immune memory, and ultimately promotes leukemia eradication. We further show that both hematologic cancers and solid tumors, including sarcomas and carcinomas, are amenable to myeloid-lineage reprogramming into TR-APCs. Finally, we demonstrate the clinical applicability of this approach by generating TR-APCs from primary clinical specimens and stimulating autologous patient-derived T cells. Thus, TR-APCs represent a cancer vaccination therapeutic strategy with broad implications for clinical immuno-oncology.Significance:Despite recent advances, the clinical benefit provided by cancer vaccination remains limited. We present a cancer vaccination approach leveraging myeloid-lineage reprogramming of cancer cells into APCs, which subsequently activate anticancer immunity through presentation of self-derived cancer antigens. Both hematologic and solid malignancies derive significant therapeutic benefit from reprogramming-based immunotherapy.This article is highlighted in the In This Issue feature, p. 1027
- Published
- 2023
11. Supplemental Table 1 from Reprogramming Cancer into Antigen-Presenting Cells as a Novel Immunotherapy
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Ravindra Majeti, Tian Yi Zhang, Robbie G. Majzner, Lindsay P. Miller, Niklas Landberg, Ian L. Linde, Christopher G. Dove, Payton L. Marshall, Melissa Stafford, Eric J. Gars, Kevin A. Nuno, Naomi L. Haddock, Feifei Zhao, Paul Phan, Sarah F. Gurev, Aaron C. Trotman-Grant, Thomas Köhnke, Amy C. Fan, and Miles H. Linde
- Abstract
Summary of clinical features of primary B-ALL specimens
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- 2023
12. Supplementary Data from NOT-Gated CD93 CAR T Cells Effectively Target AML with Minimized Endothelial Cross-Reactivity
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Crystal L. Mackall, Ravindra Majeti, Robbie G. Majzner, Ansuman T. Satpathy, Howard Y. Chang, Poul H. Sorensen, Wan-Jen Hong, Jie Liu, Htoo Zarni Oo, Mads Daugaard, Elena Sotillo, Amy Fan, Peng Xu, Kevin R. Parker, Katherine A. Freitas, Feifei Zhao, and Rebecca M. Richards
- Abstract
Supplemental figures and legends in one document
- Published
- 2023
13. Supplementary Data from Tuning the Antigen Density Requirement for CAR T-cell Activity
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Crystal L. Mackall, Ronald D. Vale, Alexander R. Dunn, Peng Xu, Rachel C. Lynn, Johanna Theruvath, Volker Wiebking, Sang M. Nguyen, Sabine Heitzeneder, Rebecca M. Richards, Aidan M. Tousley, Evan W. Weber, Meena Kadapakkam, June H. Myklebust, Louai Labanieh, Vipul T. Vachharajani, Rui Dong, Elena Sotillo, Skyler P. Rietberg, and Robbie G. Majzner
- Abstract
Supplemental Table 1 Supplementary Figures 1-10
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- 2023
14. Data from Tuning the Antigen Density Requirement for CAR T-cell Activity
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Crystal L. Mackall, Ronald D. Vale, Alexander R. Dunn, Peng Xu, Rachel C. Lynn, Johanna Theruvath, Volker Wiebking, Sang M. Nguyen, Sabine Heitzeneder, Rebecca M. Richards, Aidan M. Tousley, Evan W. Weber, Meena Kadapakkam, June H. Myklebust, Louai Labanieh, Vipul T. Vachharajani, Rui Dong, Elena Sotillo, Skyler P. Rietberg, and Robbie G. Majzner
- Abstract
Insufficient reactivity against cells with low antigen density has emerged as an important cause of chimeric antigen receptor (CAR) T-cell resistance. Little is known about factors that modulate the threshold for antigen recognition. We demonstrate that CD19 CAR activity is dependent upon antigen density and that the CAR construct in axicabtagene ciloleucel (CD19-CD28ζ) outperforms that in tisagenlecleucel (CD19-4-1BBζ) against antigen-low tumors. Enhancing signal strength by including additional immunoreceptor tyrosine-based activation motifs (ITAM) in the CAR enables recognition of low-antigen-density cells, whereas ITAM deletions blunt signal and increase the antigen density threshold. Furthermore, replacement of the CD8 hinge-transmembrane (H/T) region of a 4-1BBζ CAR with a CD28-H/T lowers the threshold for CAR reactivity despite identical signaling molecules. CARs incorporating a CD28-H/T demonstrate a more stable and efficient immunologic synapse. Precise design of CARs can tune the threshold for antigen recognition and endow 4-1BBζ-CARs with enhanced capacity to recognize antigen-low targets while retaining a superior capacity for persistence.Significance:Optimal CAR T-cell activity is dependent on antigen density, which is variable in many cancers, including lymphoma and solid tumors. CD28ζ-CARs outperform 4-1BBζ-CARs when antigen density is low. However, 4-1BBζ-CARs can be reengineered to enhance activity against low-antigen-density tumors while maintaining their unique capacity for persistence.This article is highlighted in the In This Issue feature, p. 627
- Published
- 2023
15. Long-Term Follow-up of CD19/22 CAR Therapy in Children and Young Adults with B-ALL Reveals Efficacy, Tolerability and High Survival Rates When Coupled with Hematopoietic Stem Cell Transplantation
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Liora M. Schultz, Sneha Ramakrishna, Reema Baskar, Rebecca M Richards, Jennifer Moon, Christina Baggott, Michelle Fujimoto, Michael Kunicki, Amy Li, Sneha Jariwala, Courtney Erickson, Ashley Jacobs, Karen Yamabe, Valentin Barsan, Robbie G. Majzner, Emily L. Egeler, Sharon Mavroukakis, Zachary Ehlinger, Warren D. Reynolds, Bita Sahaf, Lori Muffly, Matthew J Frank, Anne-Louise Gramstrup, Harshini Chinnasamy, Shabnum Patel, David B. Miklos, Steven A. Feldman, Crystal L. Mackall, and Kara L. Davis
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Immunology ,Cell Biology ,Hematology ,Biochemistry - Published
- 2022
16. Figure S2 from PET Reporter Gene Imaging and Ganciclovir-Mediated Ablation of Chimeric Antigen Receptor T Cells in Solid Tumors
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Sanjiv S. Gambhir, Crystal L. Mackall, Robbie G. Majzner, Jennifer R. Cochran, Dorota D. Klysz, Amin Aalipour, Elise Robinson, Corinne Beinat, Israt S. Alam, Tom Haywood, Gayatri Gowrishankar, Tara Murty, Louai Labanieh, and Surya Murty
- Abstract
Figure S2 shows characterization of B7H3-sr39tk CAR phenotypic subsets
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- 2023
17. Supplementary Data from CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors
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Crystal L. Mackall, John M. Maris, Poul H.B. Sorensen, Michelle Monje, Ravindra Majeti, Martha M. Quezado, Siddhartha S. Mitra, Ezio Bonvini, Alla Sekunova, Alberto Delaidelli, Brad St Croix, Zhongyu Zhu, Dimiter S. Dimitrov, Rimas J. Orentas, Daniel W. Lee, Louai Labanieh, Elena Sotillo, Christopher Rota, Peng Xu, Miles H. Linde, Skyler P. Rietberg, Christopher W. Mount, Yongzhi Cui, Sabine Heitzeneder, Anandani Nellan, Johanna L. Theruvath, and Robbie G. Majzner
- Abstract
Supplementary Figures
- Published
- 2023
18. Data from PET Reporter Gene Imaging and Ganciclovir-Mediated Ablation of Chimeric Antigen Receptor T Cells in Solid Tumors
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Sanjiv S. Gambhir, Crystal L. Mackall, Robbie G. Majzner, Jennifer R. Cochran, Dorota D. Klysz, Amin Aalipour, Elise Robinson, Corinne Beinat, Israt S. Alam, Tom Haywood, Gayatri Gowrishankar, Tara Murty, Louai Labanieh, and Surya Murty
- Abstract
Imaging strategies to monitor chimeric antigen receptor (CAR) T-cell biodistribution and proliferation harbor the potential to facilitate clinical translation for the treatment of both liquid and solid tumors. In addition, the potential adverse effects of CAR T cells highlight the need for mechanisms to modulate CAR T-cell activity. The herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene has previously been translated as a PET reporter gene for imaging of T-cell trafficking in patients with brain tumor. The HSV1-TK enzyme can act as a suicide gene of transduced cells through treatment with the prodrug ganciclovir. Here we report the molecular engineering, imaging, and ganciclovir-mediated destruction of B7H3 CAR T cells incorporating a mutated version of the HSV1-tk gene (sr39tk) with improved enzymatic activity for ganciclovir. The sr39tk gene did not affect B7H3 CAR T-cell functionality and in vitro and in vivo studies in osteosarcoma models showed no significant effect on B7H3 CAR T-cell antitumor activity. PET/CT imaging with 9-(4-[18F]-fluoro-3-[hydroxymethyl]butyl)guanine ([18F]FHBG) of B7H3-sr39tk CAR T cells in an orthotopic model of osteosarcoma revealed tumor homing and systemic immune expansion. Bioluminescence and PET imaging of B7H3-sr39tk CAR T cells confirmed complete tumor ablation with intraperitoneal ganciclovir administration. This imaging and suicide ablation system can provide insight into CAR T-cell migration and proliferation during clinical trials while serving as a suicide switch to limit potential toxicities.Significance:This study showcases the only genetically engineered system capable of serving the dual role both as an effective PET imaging reporter and as a suicide switch for CAR T cells.
- Published
- 2023
19. Data from CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors
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Crystal L. Mackall, John M. Maris, Poul H.B. Sorensen, Michelle Monje, Ravindra Majeti, Martha M. Quezado, Siddhartha S. Mitra, Ezio Bonvini, Alla Sekunova, Alberto Delaidelli, Brad St Croix, Zhongyu Zhu, Dimiter S. Dimitrov, Rimas J. Orentas, Daniel W. Lee, Louai Labanieh, Elena Sotillo, Christopher Rota, Peng Xu, Miles H. Linde, Skyler P. Rietberg, Christopher W. Mount, Yongzhi Cui, Sabine Heitzeneder, Anandani Nellan, Johanna L. Theruvath, and Robbie G. Majzner
- Abstract
Purpose:Patients with relapsed pediatric solid tumors and CNS malignancies have few therapeutic options and frequently die of their disease. Chimeric antigen receptor (CAR) T cells have shown tremendous success in treating relapsed pediatric acute lymphoblastic leukemia, but this has not yet translated to treating solid tumors. This is partially due to a paucity of differentially expressed cell surface molecules on solid tumors that can be safely targeted. Here, we present B7-H3 (CD276) as a putative target for CAR T-cell therapy of pediatric solid tumors, including those arising in the central nervous system.Experimental Design:We developed a novel B7-H3 CAR whose binder is derived from a mAb that has been shown to preferentially bind tumor tissues and has been safely used in humans in early-phase clinical trials. We tested B7-H3 CAR T cells in a variety of pediatric cancer models.Results:B7-H3 CAR T cells mediate significant antitumor activity in vivo, causing regression of established solid tumors in xenograft models including osteosarcoma, medulloblastoma, and Ewing sarcoma. We demonstrate that B7-H3 CAR T-cell efficacy is largely dependent upon high surface target antigen density on tumor tissues and that activity is greatly diminished against target cells that express low levels of antigen, thus providing a possible therapeutic window despite low-level normal tissue expression of B7-H3.Conclusions:B7-H3 CAR T cells could represent an exciting therapeutic option for patients with certain lethal relapsed or refractory pediatric malignancies, and should be tested in carefully designed clinical trials.
- Published
- 2023
20. Overcoming on-target, off-tumour toxicity of CAR T cell therapy for solid tumours
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Christian L. Flugel, Robbie G. Majzner, Giedre Krenciute, Gianpietro Dotti, Stanley R. Riddell, Dimitrios L. Wagner, and Mohamed Abou-el-Enein
- Subjects
Oncology ,Article - Abstract
Therapies with genetically modified T cells that express chimeric antigen receptors (CARs) specific for CD19 or B cell maturation antigen (BCMA) are approved to treat certain B cell malignancies. However, translating these successes into treatments for patients with solid tumours presents various challenges, including the risk of clinically serious on-target, off-tumour toxicity (OTOT) owing to CAR T cell-mediated cytotoxicity against non-malignant tissues expressing the target antigen. Indeed, severe OTOT has been observed in various CAR T cell clinical trials involving patients with solid tumours, highlighting the importance of establishing strategies to predict, mitigate and control the onset of this effect. In this Review, we summarize current clinical evidence of OTOT with CAR T cells in the treatment of solid tumours and discuss the utility of preclinical mouse models in predicting clinical OTOT. We then describe novel strategies being developed to improve the specificity of CAR T cells in solid tumours, particularly the role of affinity tuning of target binders, logic circuits and synthetic biology. Furthermore, we highlight control strategies that can be used to mitigate clinical OTOT following cell infusion such as regulating or eliminating CAR T cell activity, exogenous control of CAR expression, and local administration of CAR T cells.
- Published
- 2022
21. Enhanced T cell effector activity by targeting the Mediator kinase module
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Katherine A. Freitas, Julia A. Belk, Elena Sotillo, Patrick J. Quinn, Maria C. Ramello, Meena Malipatlolla, Bence Daniel, Katalin Sandor, Dorota Klysz, Jeremy Bjelajac, Peng Xu, Kylie A. Burdsall, Victor Tieu, Vandon T. Duong, Micah G. Donovan, Evan W. Weber, Howard Y. Chang, Robbie G. Majzner, Joaquin M. Espinosa, Ansuman T. Satpathy, and Crystal L. Mackall
- Subjects
Multidisciplinary ,Mediator Complex ,Receptors, Chimeric Antigen ,Cyclin C ,T-Lymphocytes ,Neoplasms ,Humans ,Genetic Testing ,Cyclin-Dependent Kinase 8 ,Immunotherapy, Adoptive ,Cyclin-Dependent Kinases ,Transcription Factors ,Genome-Wide Association Study - Abstract
T cells are the major arm of the immune system responsible for controlling and regressing cancers. To identify genes limiting T cell function, we conducted genome-wide CRISPR knockout screens in human chimeric antigen receptor (CAR) T cells. Top hits were MED12 and CCNC , components of the Mediator kinase module. Targeted MED12 deletion enhanced antitumor activity and sustained the effector phenotype in CAR- and T cell receptor–engineered T cells, and inhibition of CDK8/19 kinase activity increased expansion of nonengineered T cells. MED12 -deficient T cells manifested increased core Meditator chromatin occupancy at transcriptionally active enhancers—most notably for STAT and AP-1 transcription factors—and increased IL2RA expression and interleukin-2 sensitivity. These results implicate Mediator in T cell effector programming and identify the kinase module as a target for enhancing potency of antitumor T cell responses.
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- 2022
22. Enhanced Effector Activity of Mediator Kinase Module Deficient CAR-T Cells
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Katherine A. Freitas, Julia A. Belk, Elena Sotillo, Bence Daniel, Katalin Sandor, Dorota Klysz, Vandon T. Duong, Kylie Burdsall, Peng Xu, Meena Malipatlolla, Micah G. Donovan, Evan W. Weber, Robbie G. Majzner, Howard Y. Chang, Joaquin M. Espinosa, Ansuman T. Satpathy, and Crystal L. Mackall
- Abstract
Adoptive T cell immune therapies mediate impressive clinical benefit in a fraction of patients, but anti-tumor effects are often limited by inadequate T cell potency. To identify genes limiting T cell effector function, we conducted genome-wide CRISPR knock-out screens in human primary CAR-T cells. The top hits were MED12 and CCNC, components of the cyclin-dependent kinase (CDK) module of the Mediator complex, an evolutionarily conserved regulator of gene transcription. MED12 or CCNC deficient CAR-T cells manifest increased expansion, cytokine production, metabolic fitness, effector function, anti-tumor activity and reduced terminal effector differentiation. Chemical inhibition of CDK8/19 kinase activity recapitulated some features of genetic loss of MED12, including increased T cell expansion. MED12 deficient CAR-T cells showed widespread but selective increases in chromatin accessibility, MED1 chromatin occupancy, and H3K27 acetylation at enhancers used by transcription factors playing a critical role in T cell fate, including several STAT and AP1 family members. The most pronounced enhancement was observed for STAT5 which manifested as increased sensitivity to IL-2 in MED12 deficient T cells. These results link Mediator induced transcriptional coactivation with T cell effector programming and identify the CDK module as a target for enhancing the potency of anti-tumor T cell responses.One Sentence SummaryThe Mediator kinase module is a primary regulator of T cell differentiation, and genetic or small molecule-based inhibition of this module enhances effector T cell potency.
- Published
- 2022
23. Latent human herpesvirus 6 is reactivated in chimeric antigen receptor T cells
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Caleb A. Lareau, Yajie Yin, Katie Maurer, Katalin D. Sandor, Garima Yagnik, José Peña, Jeremy Chase Crawford, Anne M. Spanjaart, Jacob C. Gutierrez, Nicholas J. Haradhvala, Tsion Abay, Robert R. Stickels, Jeffrey M. Verboon, Vincent Liu, Jackson Southard, Ren Song, Wenjing Li, Aastha Shrestha, Laxmi Parida, Gad Getz, Marcela V. Maus, Shuqiang Li, Alison Moore, Rafael G. Amado, Aimee C. Talleur, Paul G. Thomas, Houman Dehghani, Thomas Pertel, Anshul Kundaje, Stephen Gottschalk, Theodore L. Roth, Marie J. Kersten, Catherine J. Wu, Robbie G. Majzner, and Ansuman T. Satpathy
- Abstract
Cell therapies have yielded durable clinical benefits for patients with cancer, but the risks associated with the development of therapies from manipulated human cells are still being understood. For example, we currently lack a comprehensive understanding of the mechanisms of neurotoxicity observed in patients receiving T cell therapies, including recent reports of encephalitis caused by human herpesvirus 6 (HHV-6) reactivation1. Here, via petabase-scale viral RNA data mining, we examine the landscape of human latent viral reactivation and demonstrate that HHV-6B can become reactivated in human CD4+ T cells in standard in vitro cultures. Using single-cell sequencing, we identify a rare population of HHV-6 ‘super-expressors’ (~1 in 300-10,000 cells) that possess high viral transcriptional activity in chimeric antigen receptor (CAR) T cell culture before spreading to infect other cells in vitro. Through the analysis of single-cell sequencing data from patients receiving cell therapy products that are FDA-approved2 or used in clinical studies3,4, we identify the presence of CAR+, HHV-6 super-expressor T cells in vivo. Together, our study implicates cell therapy products as a source of lytic HHV-6 reported in clinical trials1,5–7 and has broad implications for the design, production, and monitoring of cell therapies.
- Published
- 2022
24. Immunotherapy of Neuroblastoma: Facts and Hopes
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John Anderson, Robbie G. Majzner, and Paul M. Sondel
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Cancer Research ,Neuroblastoma ,Oncology ,Gangliosides ,Antibodies, Monoclonal ,Humans ,Immunologic Factors ,Immunotherapy ,Neoplasm Recurrence, Local ,Child ,Article - Abstract
While the adoption of multimodal therapy including surgery, radiation, and aggressive combination chemotherapy has improved outcomes for many children with high-risk neuroblastoma, we appear to have reached a plateau in what can be achieved with cytotoxic therapies alone. Most children with cancer, including high-risk neuroblastoma, do not benefit from treatment with immune checkpoint inhibitors (ICI) that have revolutionized the treatment of many highly immunogenic adult solid tumors. This likely reflects the low tumor mutation burden as well as the downregulated MHC-I that characterizes most high-risk neuroblastomas. For these reasons, neuroblastoma represents an immunotherapeutic challenge that may be a model for the creation of effective immunotherapy for other “cold” tumors in children and adults that do not respond to ICI. The identification of strong expression of the disialoganglioside GD2 on the surface of nearly all neuroblastoma cells provided a target for immune recognition by anti-GD2 mAbs that recruit Fc receptor–expressing innate immune cells that mediate cytotoxicity or phagocytosis. Adoption of anti-GD2 antibodies into both upfront and relapse treatment protocols has dramatically increased survival rates and altered the landscape for children with high-risk neuroblastoma. This review describes how these approaches have been expanded to additional combinations and forms of immunotherapy that have already demonstrated clear clinical benefit. We also describe the efforts to identify additional immune targets for neuroblastoma. Finally, we summarize newer approaches being pursued that may well help both innate and adaptive immune cells, endogenous or genetically engineered, to more effectively destroy neuroblastoma cells, to better induce complete remission and prevent recurrence.
- Published
- 2022
25. How to stop using gadolinium chelates for magnetic resonance imaging: clinical-translational experiences with ferumoxytol
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Michael, Heike E. Daldrup-Link, Ali Rashidi, Stuart B. Goodman, Robbie G. Majzner, Sheri L. Spunt, Michael E. Moseley, and Ashok J Theruvath
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Biodistribution ,medicine.medical_specialty ,medicine.diagnostic_test ,business.industry ,Gadolinium ,Rare earth ,chemistry.chemical_element ,Magnetic resonance imaging ,030218 nuclear medicine & medical imaging ,Food and drug administration ,Ferumoxytol ,03 medical and health sciences ,0302 clinical medicine ,chemistry ,030225 pediatrics ,Pediatrics, Perinatology and Child Health ,medicine ,Radiology, Nuclear Medicine and imaging ,Chelation ,Radiology ,business ,Neuroradiology - Abstract
Gadolinium chelates have been used as standard contrast agents for clinical MRI for several decades. However, several investigators recently reported that rare Earth metals such as gadolinium are deposited in the brain for months or years. This is particularly concerning for children, whose developing brain is more vulnerable to exogenous toxins compared to adults. Therefore, a search is under way for alternative MR imaging biomarkers. The United States Food and Drug Administration (FDA)-approved iron supplement ferumoxytol can solve this unmet clinical need: ferumoxytol consists of iron oxide nanoparticles that can be detected with MRI and provide significant T1- and T2-signal enhancement of vessels and soft tissues. Several investigators including our research group have started to use ferumoxytol off-label as a new contrast agent for MRI. This article reviews the existing literature on the biodistribution of ferumoxytol in children and compares the diagnostic accuracy of ferumoxytol- and gadolinium-chelate-enhanced MRI. Iron oxide nanoparticles represent a promising new class of contrast agents for pediatric MRI that can be metabolized and are not deposited in the brain.
- Published
- 2021
26. Coopting T cell proximal signaling molecules enables Boolean logic-gated CAR T cell control
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Aidan M. Tousley, Maria Caterina Rotiroti, Louai Labanieh, Lea Wenting Rysavy, Skyler P. Rietberg, Eva L. de la Serna, Guillermo Nicolas Dalton, Dorota Klysz, Evan W. Weber, Won-Ju Kim, Peng Xu, Elena Sotillo, Alexander R. Dunn, Crystal L. Mackall, and Robbie G. Majzner
- Abstract
Introductory paragraphWhile CAR T cells have altered the treatment landscape for B cell malignancies, the risk of on-target, off-tumor toxicity has hampered their development for solid tumors because most target antigens are shared with normal cells1,2. Researchers have attempted to apply Boolean logic gating to CAR T cells to prevent on-target, off-tumor toxicity3–7; however, a truly safe and effective logic-gated CAR has remained elusive8. Here, we describe a novel approach to CAR engineering in which we replace traditional ITAM-containing CD3ζ domains with intracellular proximal T cell signaling molecules. We demonstrate that certain proximal signaling CARs, such as a ZAP-70 CAR, can activate T cells and eradicate tumorsin vivowhile bypassing upstream signaling proteins such as CD3ζ. The primary role of ZAP-70 is to phosphorylate LAT and SLP-76, which form a scaffold for the propagation of T cell signaling. We leveraged the cooperative role of LAT and SLP-76 to engineerLogic-gatedIntracellularNetworK(LINK) CAR, a rapid and reversible Boolean-logic AND-gated CAR T cell platform that outperforms other systems in both efficacy and the prevention of on-target, off-tumor toxicity. LINK CAR will dramatically expand the number and types of molecules that can be targeted with CAR T cells, enabling the deployment of these powerful therapeutics for solid tumors and diverse diseases such as autoimmunity9and fibrosis10. In addition, this work demonstrates that the internal signaling machinery of cells can be repurposed into surface receptors, a finding that could have broad implications for new avenues of cellular engineering.
- Published
- 2022
27. Anti-GD2 synergizes with CD47 blockade to mediate tumor eradication
- Author
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Johanna Theruvath, Marie Menard, Benjamin A. H. Smith, Miles H. Linde, Garry L. Coles, Guillermo Nicolas Dalton, Wei Wu, Louise Kiru, Alberto Delaidelli, Elena Sotillo, John L. Silberstein, Anna C. Geraghty, Allison Banuelos, Molly Thomas Radosevich, Shaurya Dhingra, Sabine Heitzeneder, Aidan Tousley, John Lattin, Peng Xu, Jing Huang, Nicole Nasholm, Andy He, Tracy C. Kuo, Emma R. B. Sangalang, Jaume Pons, Amira Barkal, Rachel E. Brewer, Kristopher D. Marjon, Jose G. Vilches-Moure, Payton L. Marshall, Ricardo Fernandes, Michelle Monje, Jennifer R. Cochran, Poul H. Sorensen, Heike E. Daldrup-Link, Irving L. Weissman, Julien Sage, Ravindra Majeti, Carolyn R. Bertozzi, William A. Weiss, Crystal L. Mackall, and Robbie G. Majzner
- Subjects
Pediatric Research Initiative ,Pediatric Cancer ,Immunology ,Bone Neoplasms ,CD47 Antigen ,Medical and Health Sciences ,General Biochemistry, Genetics and Molecular Biology ,Article ,Cell Line ,Mice ,Neuroblastoma ,Rare Diseases ,Phagocytosis ,Cell Line, Tumor ,Tumor Microenvironment ,Animals ,Humans ,Cancer ,Pediatric ,Tumor ,Neurosciences ,General Medicine ,Neoplasm Recurrence ,Orphan Drug ,Local ,Immunotherapy ,Neoplasm Recurrence, Local - Abstract
The disialoganglioside GD2 is overexpressed on several solid tumors, and monoclonal antibodies targeting GD2 have substantially improved outcomes for children with high-risk neuroblastoma. However, approximately 40% of patients with neuroblastoma still relapse, and anti-GD2 has not mediated significant clinical activity in any other GD2(+) malignancy. Macrophages are important mediators of anti-tumor immunity, but tumors resist macrophage phagocytosis through expression of the checkpoint molecule CD47, a so-called ‘Don’t eat me’ signal. In this study, we establish potent synergy for the combination of anti-GD2 and anti-CD47 in syngeneic and xenograft mouse models of neuroblastoma, where the combination eradicates tumors, as well as osteosarcoma and small-cell lung cancer, where the combination significantly reduces tumor burden and extends survival. This synergy is driven by two GD2-specific factors that reorient the balance of macrophage activity. Ligation of GD2 on tumor cells (a) causes upregulation of surface calreticulin, a pro-phagocytic ‘Eat me’ signal that primes cells for removal and (b) interrupts the interaction of GD2 with its newly identified ligand, the inhibitory immunoreceptor Siglec-7. This work credentials the combination of anti-GD2 and anti-CD47 for clinical translation and suggests that CD47 blockade will be most efficacious in combination with monoclonal antibodies that alter additional pro- and anti-phagocytic signals within the tumor microenvironment.
- Published
- 2022
28. Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals
- Author
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Mary P. Hall, Mark A. Kay, Yunhee Park, Joel R. Walker, Lan Xiang Liu, Louai Labanieh, Robbie G. Majzner, Namdoo Kim, Michael Z. Lin, Lance P. Encell, Jennifer R. Cochran, Kerriann M. Casey, David C. Wang, Robin Hurst, Yichi Su, Thomas A. Kirkland, Feijie Zhang, Crystal L. Mackall, and Thomas P. Smith
- Subjects
education.field_of_study ,Tumor size ,Chemistry ,Population ,Cell Biology ,Biochemistry ,Substrate Specificity ,Luminescent Proteins ,In vivo ,Luminescent Measurements ,Biophysics ,Animals ,Substrate specificity ,Bioluminescence ,Fluorescent protein ,Bioluminescence imaging ,Luciferase ,Furans ,Luciferases ,education ,Molecular Biology ,Enzyme Assays ,Biotechnology - Abstract
Sensitive detection of two biological events in vivo has long been a goal in bioluminescence imaging. Antares, a fusion of the luciferase NanoLuc to the orange fluorescent protein CyOFP, has emerged as a bright bioluminescent reporter with orthogonal substrate specificity to firefly luciferase (FLuc) and its derivatives such as AkaLuc. However, the brightness of Antares in mice is limited by the poor solubility and bioavailability of the NanoLuc substrate furimazine. Here, we report a new substrate, hydrofurimazine, whose enhanced aqueous solubility allows delivery of higher doses to mice. In the liver, Antares with hydrofurimazine exhibited similar brightness to AkaLuc with its substrate AkaLumine. Further chemical exploration generated a second substrate, fluorofurimazine, with even higher brightness in vivo. We used Antares with fluorofurimazine to track tumor size and AkaLuc with AkaLumine to visualize CAR-T cells within the same mice, demonstrating the ability to perform two-population imaging with these two luciferase systems. NanoLuc substrates with improved solubility and bioavailability, hydrofurimazine and fluorofurimazine, strongly enhance bioluminescence signals in vivo and enable bright dual-color bioluminescent imaging with AkaLuc and AkaLumine.
- Published
- 2020
29. Immune-Based Approaches for the Treatment of Pediatric Malignancies
- Author
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Kristopher R. Bosse, Crystal L. Mackall, Robbie G. Majzner, and John M. Maris
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,Article ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,pediatric cancers ,B-cell acute lymphocytic leukemia ,antibodies ,Medicine ,CAR T cells ,biology ,business.industry ,Cell Biology ,Immunotherapy ,Chimeric antigen receptor ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,immunotherapy ,Antibody ,Car t cells ,business - Abstract
Immune-based therapies have now been credentialed for pediatric cancers with the robust efficacy of chimeric antigen receptor (CAR) T cells for pediatric B cell acute lymphocytic leukemia (ALL), offering a chance of a cure for children with previously lethal disease and a potentially more targeted therapy to limit treatment-related morbidities. The developmental origins of most pediatric cancers make them ideal targets for immune-based therapies that capitalize on the differential expression of lineage-specific cell surface molecules such as antibodies, antibody-drug conjugates, or CAR T cells, while the efficacy of other therapies that depend on tumor immunogenicity such as immune checkpoint inhibitors has been limited to date. Here we review the current status of immune-based therapies for childhood cancers, discuss challenges to developing immunotherapeutics for these diseases, and outline future directions of pediatric immunotherapy discovery and development.
- Published
- 2020
30. c-Jun overexpression in CAR T cells induces exhaustion resistance
- Author
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Robert C. Jones, Surya Nagaraja, Ansuman T. Satpathy, Michelle Monje, Zinaida Good, Elena Sotillo, Howard Y. Chang, Peng Xu, Hima Anbunathan, Evan W. Weber, Crystal L. Mackall, Rachel C. Lynn, Charles F. A. de Bourcy, David Gennert, John Lattin, Jeffrey M. Granja, Robbie G. Majzner, Victor Tieu, and Stephen R. Quake
- Subjects
0301 basic medicine ,Transcription, Genetic ,Proto-Oncogene Proteins c-jun ,T cell ,T-Lymphocytes ,Receptors, Antigen, T-Cell ,Article ,Epigenesis, Genetic ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Antigen ,Cell Line, Tumor ,Neoplasms ,medicine ,Animals ,Humans ,Receptor ,Transcription factor ,Multidisciplinary ,Chemistry ,c-jun ,Chimeric antigen receptor ,Cell biology ,Chromatin ,Transcription Factor AP-1 ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Cell culture ,030220 oncology & carcinogenesis ,human activities - Abstract
Chimeric antigen receptor (CAR) T cells mediate anti-tumour effects in a small subset of patients with cancer1-3, but dysfunction due to T cell exhaustion is an important barrier to progress4-6. To investigate the biology of exhaustion in human T cells expressing CAR receptors, we used a model system with a tonically signaling CAR, which induces hallmark features of exhaustion6. Exhaustion was associated with a profound defect in the production of IL-2, along with increased chromatin accessibility of AP-1 transcription factor motifs and overexpression of the bZIP and IRF transcription factors that have been implicated in mediating dysfunction in exhausted T cells7-10. Here we show that CAR T cells engineered to overexpress the canonical AP-1 factor c-Jun have enhanced expansion potential, increased functional capacity, diminished terminal differentiation and improved anti-tumour potency in five different mouse tumour models in vivo. We conclude that a functional deficiency in c-Jun mediates dysfunction in exhausted human T cells, and that engineering CAR T cells to overexpress c-Jun renders them resistant to exhaustion, thereby addressing a major barrier to progress for this emerging class of therapeutic agents.
- Published
- 2019
31. Determinants of resistance to engineered T cell therapies targeting CD19 in large B cell lymphomas
- Author
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Brian J. Sworder, David M. Kurtz, Stefan K. Alig, Matthew J. Frank, Navika Shukla, Andrea Garofalo, Charles W. Macaulay, Mohammad Shahrokh Esfahani, Mari N. Olsen, James Hamilton, Hitomi Hosoya, Mark Hamilton, Jay Y. Spiegel, John H. Baird, Takeshi Sugio, Mia Carleton, Alexander F.M. Craig, Sheren F. Younes, Bita Sahaf, Natasha D. Sheybani, Joseph G. Schroers-Martin, Chih Long Liu, Jean S. Oak, Michael C. Jin, Sara Beygi, Andreas Hüttmann, Christine Hanoun, Ulrich Dührsen, Jason R. Westin, Michael S. Khodadoust, Yasodha Natkunam, Robbie G. Majzner, Crystal L. Mackall, Maximilian Diehn, David B. Miklos, and Ash A. Alizadeh
- Subjects
Cancer Research ,Oncology ,Medizin - Abstract
Most relapsed/refractory large B cell lymphoma (r/rLBCL) patients receiving anti-CD19 chimeric antigen receptor (CAR19) T cells relapse. To characterize determinants of resistance, we profiled over 700 longitudinal specimens from two independent cohorts (n = 65 and n = 73) of r/rLBCL patients treated with axicabtagene ciloleucel. A method for simultaneous profiling of circulating tumor DNA (ctDNA), cell-free CAR19 (cfCAR19) retroviral fragments, and cell-free T cell receptor rearrangements (cfTCR) enabled integration of tumor and both engineered and non-engineered T cell effector-mediated factors for assessing treatment failure and predicting outcomes. Alterations in multiple classes of genes are associated with resistance, including B cell identity (PAX5 and IRF8), immune checkpoints (CD274), and those affecting the microenvironment (TMEM30A). Somatic tumor alterations affect CAR19 therapy at multiple levels, including CAR19 T cell expansion, persistence, and tumor microenvironment. Further, CAR19 T cells play a reciprocal role in shaping tumor genotype and phenotype. We envision these findings will facilitate improved chimeric antigen receptor (CAR) T cells and personalized therapeutic approaches.
- Published
- 2021
32. Abstract PR003: Lineage plasticity dictates responsiveness to anti-GD2 therapy in neuroblastoma
- Author
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Nathaniel W. Mabe, Min Huang, Daniel A. Schaefer, Guillermo N. Dalton, Giulia Digiovanni, Gabriela Alexe, Anna C. Geraghty, Delan Khalid, Marius M. Mader, Michal Sheffer, Miles H. Linde, Nghi Ly, Maria Caterina Rotiroti, Benjamin A. H. Smith, Marius Wernig, Carolyn R. Bertozzi, Michelle Monje, Constantine Mitsiades, Ravindra Majeti, Ansuman T. Satpathy, Kimberly Stegmaier, and Robbie G. Majzner
- Subjects
Cancer Research ,Oncology - Abstract
Epigenetic dysregulation is frequently observed in the disease pathology of pediatric cancers, including neuroblastoma, the most common extracranial solid tumor in pediatric patients. Neuroblastoma tumors co-opt developmentally linked adrenergic or mesenchymal super-enhancer landscapes that rewire their transcriptional programs. Here, we describe that the lineage commitment to a mesenchymal epigenetic state is an important mechanism of resistance to anti-GD2 therapy through loss of GD2 antigen, a ganglioside glycolipid expressed on the cell surface. Low GD2 expression was significantly correlated with the mesenchymal state in a large panel of neuroblastoma cell lines and a forced adrenergic-to-mesenchymal transition conferred downregulation of GD2 and resistance to anti-GD2 antibody. Mechanistically, low-GD2 expressing cell lines demonstrated significantly reduced expression of the ganglioside synthesis enzyme ST8SIA1 (GD3 synthase), resulting in a bottlenecking of GD2 synthesis. Genome-wide CRISPR/Cas9 screening to identify regulators of GD2 in neuroblastoma revealed that the ablation of the polycomb repressive complex 2 (PRC2) significantly upregulates GD2 expression in GD2-low cells. Pharmacologic inhibition of EZH2 resulted in epigenetic rewiring of mesenchymal neuroblastoma cells into an adrenergic-like state, re-expressed ST8SIA1, and restored surface expression of GD2 and sensitivity to anti-GD2 antibody. These data identify developmental lineage as a key determinant of sensitivity to anti-GD2 based immunotherapies and credential PRC2 inhibitors for clinical testing in combination with anti-GD2 antibody to enhance outcomes for children with neuroblastoma. Citation Format: Nathaniel W. Mabe, Min Huang, Daniel A. Schaefer, Guillermo N. Dalton, Giulia Digiovanni, Gabriela Alexe, Anna C. Geraghty, Delan Khalid, Marius M. Mader, Michal Sheffer, Miles H. Linde, Nghi Ly, Maria Caterina Rotiroti, Benjamin A. H. Smith, Marius Wernig, Carolyn R. Bertozzi, Michelle Monje, Constantine Mitsiades, Ravindra Majeti, Ansuman T. Satpathy, Kimberly Stegmaier, Robbie G. Majzner. Lineage plasticity dictates responsiveness to anti-GD2 therapy in neuroblastoma. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr PR003.
- Published
- 2022
33. DETERMINANTS OF RESISTANCE TO ENGINEERED T‐CELL THERAPIES TARGETING CD19 IN LYMPHOMA
- Author
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Jay Y. Spiegel, Matthew J. Frank, Stefan Alig, N Sheybani, Sara Beygi, Mari Olsen, Michael S. Khodadoust, Joseph G Schroers-Martin, David M. Kurtz, Yasodha Natkunam, Navika D. Shukla, Robbie G. Majzner, Bita Sahaf, Brian Sworder, Michael C. Jin, D. M Miklos, Jean Oak, C. W Macauley, Maximilian Diehn, Andrea Garofalo, Ash A. Alizadeh, C. L. Liu, Mohammad Shahrokh Esfahani, and Crystal L. Mackall
- Subjects
Cancer Research ,biology ,business.industry ,T cell ,Hematology ,General Medicine ,medicine.disease ,CD19 ,Lymphoma ,medicine.anatomical_structure ,Oncology ,medicine ,biology.protein ,Cancer research ,business - Published
- 2021
34. Charting a path for prioritization of novel agents for clinical trials in osteosarcoma: A report from the Children's Oncology Group New Agents for Osteosarcoma Task Force
- Author
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Fariba Navid, Ryan D. Roberts, Damon R. Reed, Sarah B. Whittle, Katherine A. Janeway, Katharine Offer, Scott C. Borinstein, Cheryl A. London, Alex Yee-Chen Huang, Robbie G. Majzner, Michael W. Bishop, Emily K. Slotkin, Pooja Hingorani, Patrick J. Grohar, Michael S. Isakoff, Emily Greengard, Richard Gorlick, Peter J. Houghton, Elizabeth Stewart, E. Alejandro Sweet Cordero, and Amy K. LeBlanc
- Subjects
Oncology ,Prioritization ,medicine.medical_specialty ,Bone Neoplasms ,Disease ,Article ,Metastasis ,Epigenesis, Genetic ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,Internal medicine ,medicine ,Humans ,Child ,Protein Kinase Inhibitors ,Clinical Trials as Topic ,Osteosarcoma ,business.industry ,Task force ,Hematology ,RELAPSED DISEASE ,medicine.disease ,Clinical trial ,Novel agents ,030220 oncology & carcinogenesis ,Pediatrics, Perinatology and Child Health ,Immunotherapy ,business ,030215 immunology - Abstract
Osteosarcoma is the most common bone tumor in children and young adults. Metastatic and relapsed disease confer poor prognosis, and there have been no improvements in outcomes for several decades. The disease’s biological complexity, lack of drugs developed specifically for osteosarcoma, imperfect pre-clinical models, and limits of existing clinical trial designs have contributed to lack of progress. The Children’s Oncology Group Bone Tumor Committee established the New Agents for Osteosarcoma Task Force to identify and prioritize agents for inclusion in clinical trials. The group identified multi-targeted tyrosine kinase inhibitors, immunotherapies targeting B7-H3, CD47-SIRPα inhibitors, telaglenastat, and epigenetic modifiers as the top agents of interest. Only multi-targeted tyrosine kinase inhibitors met all criteria for front-line evaluation and have already been incorporated into an upcoming phase III study concept. The task force will continue to reassess identified agents of interest as new data becomes available and evaluate novel agents using this method.
- Published
- 2021
35. Clinical lessons learned from the first leg of the CAR T cell journey
- Author
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Robbie G. Majzner and Crystal L. Mackall
- Subjects
0301 basic medicine ,Oncology ,medicine.medical_specialty ,T cell ,Cell ,Cancer ,General Medicine ,Disease ,medicine.disease ,General Biochemistry, Genetics and Molecular Biology ,Chimeric antigen receptor ,Clinical trial ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Internal medicine ,medicine ,Car t cells ,B cell - Abstract
Chimeric antigen receptor (CAR) T cell therapy for B cell malignancies has surpassed expectations, driving an ever-expanding number of clinical trials and the first US Food and Drug Administration approvals of cell therapies for the treatment of cancer. This experience has illuminated some generalizable requirements for CAR T cell efficacy as well as the interplay between disease biology and clinical outcomes. Major CAR intrinsic variables affecting T cell behavior have been defined, and mechanisms of tumor resistance are increasingly understood. Here, we review the clinical experience with CAR T cells amassed to date, including but not limited to B cell malignancies, emphasizing factors associated with efficacy, resistance and major barriers to success. We also discuss how these insights are driving next-generation clinical trials, including those in solid tumors.
- Published
- 2019
36. CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors
- Author
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Siddhartha Mitra, Zhongyu Zhu, Anandani Nellan, Johanna Theruvath, Ezio Bonvini, Skyler P. Rietberg, Ravindra Majeti, Christopher Rota, Sabine Heitzeneder, Crystal L. Mackall, Elena Sotillo, Yongzhi Cui, Peng Xu, Alla Sekunova, Christopher Mount, Alberto Delaidelli, John M. Maris, Daniel W. Lee, Michelle Monje, Brad St. Croix, Robbie G. Majzner, Dimiter S. Dimitrov, Miles H. Linde, Poul H. Sorensen, Martha Quezado, Louai Labanieh, and Rimas J. Orentas
- Subjects
0301 basic medicine ,Cancer Research ,B7 Antigens ,T-Lymphocytes ,medicine.medical_treatment ,Receptors, Antigen, T-Cell ,Immunotherapy, Adoptive ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,Antigens, Neoplasm ,Cell Line, Tumor ,Animals ,Humans ,Medicine ,Medulloblastoma ,Receptors, Chimeric Antigen ,Brain Neoplasms ,business.industry ,Immunotherapy ,medicine.disease ,Immunohistochemistry ,Xenograft Model Antitumor Assays ,Pediatric cancer ,Chimeric antigen receptor ,Disease Models, Animal ,Treatment Outcome ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Osteosarcoma ,Sarcoma ,business - Abstract
Purpose: Patients with relapsed pediatric solid tumors and CNS malignancies have few therapeutic options and frequently die of their disease. Chimeric antigen receptor (CAR) T cells have shown tremendous success in treating relapsed pediatric acute lymphoblastic leukemia, but this has not yet translated to treating solid tumors. This is partially due to a paucity of differentially expressed cell surface molecules on solid tumors that can be safely targeted. Here, we present B7-H3 (CD276) as a putative target for CAR T-cell therapy of pediatric solid tumors, including those arising in the central nervous system. Experimental Design: We developed a novel B7-H3 CAR whose binder is derived from a mAb that has been shown to preferentially bind tumor tissues and has been safely used in humans in early-phase clinical trials. We tested B7-H3 CAR T cells in a variety of pediatric cancer models. Results: B7-H3 CAR T cells mediate significant antitumor activity in vivo, causing regression of established solid tumors in xenograft models including osteosarcoma, medulloblastoma, and Ewing sarcoma. We demonstrate that B7-H3 CAR T-cell efficacy is largely dependent upon high surface target antigen density on tumor tissues and that activity is greatly diminished against target cells that express low levels of antigen, thus providing a possible therapeutic window despite low-level normal tissue expression of B7-H3. Conclusions: B7-H3 CAR T cells could represent an exciting therapeutic option for patients with certain lethal relapsed or refractory pediatric malignancies, and should be tested in carefully designed clinical trials.
- Published
- 2019
37. Abstract 2822: Enhanced effector activity of mediator CDK8 kinase module deficient CAR-T Cells
- Author
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Katherine A. Freitas, Julia A. Belk, Elena Sotillo, Bence Daniel, Katalin Sandor, Dorota Klysz, Vandon T. Duong, Peng Xu, Meena Malipatlolla, Evan W. Weber, Robbie G. Majzner, Howard Y. Chang, Ansuman T. Satpathy, and Crystal Mackall
- Subjects
Cancer Research ,Oncology - Abstract
Adoptive T cell immune therapies mediate impressive clinical benefit in a fraction of patients, but anti-tumor effects are often limited by inadequate T cell potency. To identify genes that limit T cell effector function, we conducted genome-wide CRISPR knock-out screens in human primary CAR-T cells. The top hits were components of the CDK8 kinase module of the Mediator complex, an evolutionarily conserved regulator of gene transcription. CDK8 kinase module deficient CAR-T cells manifest increased expansion, cytokine production, metabolic fitness, effector function, anti-tumor activity and reduced terminal effector differentiation. CDK8 kinase module deficient CAR-T cells showed widespread but selective increases in chromatin accessibility, MED1 chromatin occupancy, and H3K27 acetylation most notably involving transcription factors that play a critical role in T cell fate, including several STAT and AP1 family members. The most pronounced enhancement was observed for STAT5 which manifested as increased sensitivity to IL-2 in CDK8 kinase module deficient CAR-T cells. These results link Mediator induced transcriptional coactivation with T cell effector programming and identify the CDK8 kinase module as a target for enhancing the potency of anti-tumor T cell responses. Citation Format: Katherine A. Freitas, Julia A. Belk, Elena Sotillo, Bence Daniel, Katalin Sandor, Dorota Klysz, Vandon T. Duong, Peng Xu, Meena Malipatlolla, Evan W. Weber, Robbie G. Majzner, Howard Y. Chang, Ansuman T. Satpathy, Crystal Mackall. Enhanced effector activity of mediator CDK8 kinase module deficient CAR-T Cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2822.
- Published
- 2022
38. Abstract CT001: Major tumor regressions in H3K27M-mutated diffuse midline glioma (DMG) following sequential intravenous (IV) and intracerebroventricular (ICV) delivery of GD2-CAR T cells
- Author
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Robbie G. Majzner, Jasia Mahdi, Sneha Ramakrishna, Shabnum Patel, Harshini Chinnasamy, Kristen Yeom, Liora Schultz, Valentin Barsan, Rebecca Richards, Cynthia Campen, Agnes Reschke, Angus Martin Shaw Toland, Christina Baggott, Sharon Mavroukakis, Emily Egeler, Jennifer Moon, Ashley Jacobs, Karen Yamabe-Kwong, Lindsey Rasmussen, Esther Nie, Sean Green, Michael Kunicki, Michelle Fujimoto, Zach Ehlinger, Warren Reynolds, Snehit Prabhu, Katherine E. Warren, Tim Cornell, Sonia Partap, Paul Fisher, Gerald Grant, Hannes Vogel, Bita Sahaf, Kara Davis, Steven Feldman, Michelle Monje, and Crystal L. Mackall
- Subjects
Cancer Research ,Oncology - Abstract
Background: H3K27M-mutated DMGs are universally lethal central nervous system tumors that express high levels of the disialoganglioside GD2. IV administered GD2-CAR T cells (GD2-CART) regress DMG in preclinical models, and locoregionally delivered CARs demonstrate enhanced activity in xenograft models of brain tumors. Methods: NCT04196413 is a 3+3 Phase I dose escalation trial testing GD2-CART in patients with H3K27M DMG, with dose-limiting toxicities (DLT) considered independently for DIPG and spinal DMG (sDMG). Arm A tested escalating doses of IV GD2-CART (DL1: 1e6 GD2-CART/kg; DL2=3e6 GD2-CART/kg) following lymphodepletion (LD). After the DLT period, patients with clinical and/or radiographic benefit were eligible for subsequent ICV GD2-CART (10-30e6 GD2-CART) administered via Ommaya catheter without LD every 4-8 weeks for a maximum of 12 doses. We previously reported early results from 4 patients treated on DL1, which demonstrated clinical activity and manageable toxicity. Here we provide updated results for DL1 and DL2. Results: Thirteen subjects were enrolled and 11 treated [n=4 DL1 (3 DIPG/1 sDMG); n=9 DL2 (7 DIPG/2 sDMG)]. Two subjects were removed prior to treatment due to rapid progression. No DLTs were observed on DL1. Three subjects experienced DLT on DL2 (2 DIPG/1 sDMG) due to grade 4 cytokine release syndrome (CRS), successfully managed with tocilizumab, anakinra, and corticosteroids. CRS occurred earlier on DL2 vs. DL1 (Day 3 vs 7). On both dose levels, all subjects exhibited transient symptoms related to on-tumor inflammation, termed Tumor Inflammation-Associated Neurotoxicity (TIAN), which was successfully managed with anakinra and, in some cases, CSF drainage and dexamethasone. No DLT due to TIAN has occurred. Ten patients have had adequate follow-up to assess benefit. Nine experienced radiographic and/or clinical benefit after IV infusion, and they received subsequent ICV GD2-CART infusions (median= 4 ICV infusions/pt, range 1-6). ICV infusions were not associated with high-grade CRS, although some subjects developed transient fever, headache, meningismus, nausea, and/or vomiting, and several subjects developed TIAN. Four patients continue to receive ICV infusions on study and have experienced continued clinical and radiographic benefit at 11+, 9.5+, 8+ and 7+ months following enrollment. A 31-year-old with sDMG has experienced a near-complete (>95%) reduction in tumor volume and a 17-year-old with DIPG experienced a near-complete (>98%) reduction in volume of a pontine tumor. Conclusions: IV treatment of DIPG and sDMG with GD2-CART is safe at a dose of 1e6/kg, but associated with unacceptable rates of high-grade CRS at 3e6/kg. ICV GD2-CART without LD, administered following a previous course of IV GD2-CART with LD, has been well tolerated and has mediated impressive sustained clinical benefit in some patients with DIPG/sDMG. Given these findings, we are launching a new arm to assess safety and activity and to define the recommended phase 2 dose for ICV delivery of GD2-CART without LD. Patients are eligible for up to 12 ICV infusions of GD2-CART administered every 4-6 weeks. Clinical benefit will be formally assessed using patient-reported outcomes. GD2-CART has the potential to transform therapy for patients with H3K27M+ DIPG/sDMG. Citation Format: Robbie G. Majzner, Jasia Mahdi, Sneha Ramakrishna, Shabnum Patel, Harshini Chinnasamy, Kristen Yeom, Liora Schultz, Valentin Barsan, Rebecca Richards, Cynthia Campen, Agnes Reschke, Angus Martin Shaw Toland, Christina Baggott, Sharon Mavroukakis, Emily Egeler, Jennifer Moon, Ashley Jacobs, Karen Yamabe-Kwong, Lindsey Rasmussen, Esther Nie, Sean Green, Michael Kunicki, Michelle Fujimoto, Zach Ehlinger, Warren Reynolds, Snehit Prabhu, Katherine E. Warren, Tim Cornell, Sonia Partap, Paul Fisher, Gerald Grant, Hannes Vogel, Bita Sahaf, Kara Davis, Steven Feldman, Michelle Monje, Crystal L. Mackall. Major tumor regressions in H3K27M-mutated diffuse midline glioma (DMG) following sequential intravenous (IV) and intracerebroventricular (ICV) delivery of GD2-CAR T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT001.
- Published
- 2022
39. Enhanced safety and efficacy of protease-regulated CAR-T cell receptors
- Author
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Louai Labanieh, Robbie G. Majzner, Dorota Klysz, Elena Sotillo, Chris J. Fisher, José G. Vilches-Moure, Kaithlen Zen B. Pacheco, Meena Malipatlolla, Peng Xu, Jessica H. Hui, Tara Murty, Johanna Theruvath, Nishant Mehta, Sean A. Yamada-Hunter, Evan W. Weber, Sabine Heitzeneder, Kevin R. Parker, Ansuman T. Satpathy, Howard Y. Chang, Michael Z. Lin, Jennifer R. Cochran, and Crystal L. Mackall
- Subjects
General Biochemistry, Genetics and Molecular Biology - Published
- 2022
40. Augmenting anti-CD19 and anti-CD22 CAR T-cell function using PD-1-CD28 checkpoint fusion proteins
- Author
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Nicola Habjan, Franziska Blaeschke, Felicitas Rataj, Bruno L Cadilha, Theresa Kaeuferle, Sebastian Kobold, Jasmin Mahdawi, Tobias Feuchtinger, Robbie G. Majzner, Antonia Apfelbeck, Dana Stenger, Eva Ortner, Mohamed-Reda Benmebarek, Mareike Lepenies, Semjon Willier, and Dirk Büsch
- Subjects
Recombinant Fusion Proteins ,Antigens, CD19 ,Programmed Cell Death 1 Receptor ,Immunotherapy, Adoptive ,Text mining ,CD28 Antigens ,Cell Line, Tumor ,Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ,Correspondence ,Medicine ,Humans ,CD22 CAR-T ,RC254-282 ,Acute lymphocytic leukaemia ,business.industry ,Receptors, IgE ,Anti cd19 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,CD28 ,Hematology ,Fusion protein ,Cell function ,Oncology ,Cancer research ,Immunotherapy ,business - Published
- 2021
41. Anti-GD2 antibody disrupts GD2:Siglec-7 interactions and synergizes with CD47 blockade to mediate tumor eradication
- Author
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Heike E. Daldrup-Link, John Lattin, Menard M, Pons J, Peng Xu, Kristopher D. Marjon, Aidan Tousley, Louise Kiru, John L. Silberstein, Alberto Delaidelli, Payton L. Marshall, Nasholm N, William A. Weiss, Allison Banuelos, Amira A. Barkal, Poul H. Sorensen, Garry L. Coles, Irving L. Weissman, Sabine Heitzeneder, Benjamin A. H. Smith, Robbie G. Majzner, Ricardo A. Fernandes, Jonathan Huang, Julien Sage, Miles H. Linde, Shaurya Dhingra, Elena Sotillo, Johanna Theruvath, Rachel E. Brewer, Ravi Majeti, Cochran, Sangalang Er, Crystal L. Mackall, Dalton Gn, Wei Wu, Carolyn R. Bertozzi, Kuo Tc, and Andy He
- Subjects
biology ,business.industry ,CD47 ,SIGLEC ,medicine.disease ,Blockade ,Neuroblastoma ,biology.protein ,Cancer research ,Macrophage ,Medicine ,Cytotoxic T cell ,Osteosarcoma ,Antibody ,business - Abstract
The disialoganglioside GD2 is consistently overexpressed in neuroblastoma and osteosarcoma, and is variably expressed in other sarcomas, gliomas, neuroendocrine tumors, and epithelial cancers. Anti-GD2 antibodies have improved the survival rates of patients with neuroblastoma only when administered as part of intense chemotherapy-based cytotoxic regimens, which are associated with debilitating late effects including hearing loss, growth retardation, and secondary leukemias. Despite broad expression of GD2 on osteosarcoma, anti-GD2 antibody has not mediated significant antitumor activity in that disease or any other GD2+ cancers. CD47 is a checkpoint molecule overexpressed on tumor cells that inhibits macrophage activity, and CD47 blockade has demonstrated promising clinical activity in early human trials. We investigated whether anti-CD47 antibody could enhance the efficacy of anti-GD2 antibody in neuroblastoma and other GD2+ malignancies. We demonstrate substantial synergy of these two agents, resulting in the recruitment of tumor associated macrophages (TAMs) to mediate robust and durable anti-tumor responses. The responses are driven by GD2-specific factors that reorient the balance of macrophage activity towards phagocytosis of tumor cells, including disruption of a newly described GD2:Siglec-7 axis. These results demonstrate the unique synergy of combining anti-GD2 with anti-CD47, which has the potential to significantly enhance outcomes for children with neuroblastoma and osteosarcoma and will soon be investigated in a first-in-human clinical trial.
- Published
- 2021
42. NOT-Gated CD93 CAR T Cells Effectively Target AML with Minimized Endothelial Cross-Reactivity
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Howard Y. Chang, Rebecca Richards, Peng Xu, Jie Liu, Robbie G. Majzner, Wan-Jen Hong, Ansuman T. Satpathy, Mads Daugaard, Ravindra Majeti, Feifei Zhao, Amy Fan, Poul H. Sorensen, Elena Sotillo, Katherine A. Freitas, Htoo Zarni Oo, Kevin R. Parker, and Crystal L. Mackall
- Subjects
T-Lymphocytes ,Myeloid leukemia ,Endothelial Cells ,Context (language use) ,General Medicine ,Biology ,Immunotherapy, Adoptive ,In the Spotlight ,Chimeric antigen receptor ,Proinflammatory cytokine ,Transcriptome ,Endothelial stem cell ,Haematopoiesis ,Leukemia, Myeloid, Acute ,Mice ,Cell Line, Tumor ,Cancer research ,Animals ,Humans ,Progenitor cell ,human activities ,Research Articles - Abstract
CD93 CAR T cells eliminate AML in preclinical models without targeting hematopoietic progenitor cells, and a NOT-gated CAR engineering strategy mitigates on-target, off-tumor toxicity to endothelial cells., Chimeric antigen receptor (CAR) T cells hold promise for the treatment of acute myeloid leukemia (AML), but optimal targets remain to be defined. We demonstrate that CD93 CAR T cells engineered from a novel humanized CD93-specific binder potently kill AML in vitro and in vivo but spare hematopoietic stem and progenitor cells (HSPC). No toxicity is seen in murine models, but CD93 is expressed on human endothelial cells, and CD93 CAR T cells recognize and kill endothelial cell lines. We identify other AML CAR T-cell targets with overlapping expression on endothelial cells, especially in the context of proinflammatory cytokines. To address the challenge of endothelial-specific cross-reactivity, we provide proof of concept for NOT-gated CD93 CAR T cells that circumvent endothelial cell toxicity in a relevant model system. We also identify candidates for combinatorial targeting by profiling the transcriptome of AML and endothelial cells at baseline and after exposure to proinflammatory cytokines. Significance: CD93 CAR T cells eliminate AML and spare HSPCs but exert on-target, off-tumor toxicity to endothelial cells. We show coexpression of other AML targets on endothelial cells, introduce a novel NOT-gated strategy to mitigate endothelial toxicity, and demonstrate use of high-dimensional transcriptomic profiling for rational design of combinatorial immunotherapies. See related commentary by Velasquez and Gottschalk, p. 559. This article is highlighted in the In This Issue feature, p. 549
- Published
- 2020
43. Immunotherapy for Pediatric Sarcomas
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Karin Straathof, Robbie G. Majzner, and Allison Pribnow
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Oncology ,medicine.medical_specialty ,business.industry ,medicine.drug_class ,medicine.medical_treatment ,Multimodal therapy ,Combination chemotherapy ,B-cell acute lymphoblastic leukemia ,Immunotherapy ,medicine.disease ,Monoclonal antibody ,Internal medicine ,Neuroblastoma ,Pediatric oncology ,Medicine ,In patient ,business - Abstract
While pediatric oncology saw great advances in patient survival throughout the second half of the twentieth century with the adoption of combination chemotherapy and multimodal therapy (Smith et al. 2014), this effect has mostly plateaued over the past two decades. This is especially noticeable in high-risk metastatic and relapsed sarcomas, where there has been little progress in improving patient outcomes for over 30 years. Therefore, the success of immunotherapy across a broad spectrum of cancers, including malignancies resistant to other forms of conventional cytotoxic chemotherapy or targeted therapies (Brahmer et al. 2015; Postow et al. 2015; Brown et al. 2016; Robert et al. 2015), has sparked an interest in harnessing these therapies to treat pediatric sarcomas. In pediatric oncology in general, immunotherapy has already altered the landscape of relapsed B cell acute lymphoblastic leukemia (anti-CD19 CAR T cells (Lee et al. 2015; Maude et al. 2014, 2018; Gardner et al. 2017) and bispecific antibodies (Gore et al. 2018; von Stackelberg et al. 2016)) and high-risk neuroblastoma (anti-GD2 monoclonal antibodies (Yu et al. 2010; Kushner et al. 2018)). To date, the efficacy of immunotherapy in the treatment of pediatric sarcomas has been disappointing, and this approach has not yet benefited large numbers of patients. However, early results from several trials indicate that this powerful strategy will eventually bear fruit, adding to the multimodal armament needed to cure high-risk malignancies.
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- 2020
44. GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity
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Sabine Heitzeneder, Kristopher R. Bosse, Zhongyu Zhu, Doncho Zhelev, Robbie G. Majzner, Molly T. Radosevich, Shaurya Dhingra, Elena Sotillo, Samantha Buongervino, Guillem Pascual-Pasto, Emily Garrigan, Peng Xu, Jing Huang, Benjamin Salzer, Alberto Delaidelli, Swetha Raman, Hong Cui, Benjamin Martinez, Scott J. Bornheimer, Bita Sahaf, Anya Alag, Irfete S. Fetahu, Martin Hasselblatt, Kevin R. Parker, Hima Anbunathan, Jennifer Hwang, Min Huang, Kathleen Sakamoto, Norman J. Lacayo, Dorota D. Klysz, Johanna Theruvath, José G. Vilches-Moure, Ansuman T. Satpathy, Howard Y. Chang, Manfred Lehner, Sabine Taschner-Mandl, Jean-Phillipe Julien, Poul H. Sorensen, Dimiter S. Dimitrov, John M. Maris, and Crystal L. Mackall
- Subjects
Neuroblastoma ,Cancer Research ,Receptors, Chimeric Antigen ,Glypicans ,Oncology ,Cell Line, Tumor ,T-Lymphocytes ,Receptors, Antigen, T-Cell ,Animals ,Humans ,Immunotherapy ,Immunotherapy, Adoptive ,Xenograft Model Antitumor Assays - Abstract
Pediatric cancers often mimic fetal tissues and express proteins normally silenced postnatally that could serve as immune targets. We developed T cells expressing chimeric antigen receptors (CARs) targeting glypican-2 (GPC2), a fetal antigen expressed on neuroblastoma (NB) and several other solid tumors. CARs engineered using standard designs control NBs with transgenic GPC2 overexpression, but not those expressing clinically relevant GPC2 site density (∼5,000 molecules/cell, range 1-6 × 10
- Published
- 2022
45. EPCT-14. GD2 CAR T-CELLS MEDIATE CLINICAL ACTIVITY AND MANAGEABLE TOXICITY IN CHILDREN AND YOUNG ADULTS WITH H3K27M-MUTATED DIPG AND SPINAL CORD DMG
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Bita Sahaf, Sreevidya Kurra, Michelle Fujimoto, Anne Cunniffe Marcy, Crystal L. Mackall, Emily Egeler, Gerald A. Grant, Angus Toland, Kayla Landrum, John S. Tamaresis, Sneha Ramakrishna, Rebecca Richards, Paul G. Fisher, Kara L. Davis, Courtney Erickson, Steven A. Feldman, Sharon Mavroukakis, Michael Kunicki, Michelle Monje, Timothy T. Cornell, Sonia Partap, Agnes Reschke, Lindsay Rasmussen, Jasia Mahdi, Valentin Barsan, Hannes Vogel, Robbie G. Majzner, Cynthia J. Campen, Jennifer Moon, Zach Ehlinger, Christina Baggott, Kristen W. Yeom, Liora M. Schultz, Harshini Chinnasamy, Shabnum Patel, and Aaron Mochizuki
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Oncology ,Cancer Research ,medicine.medical_specialty ,Cyclophosphamide ,business.industry ,Phases of clinical research ,Inflammation ,medicine.disease ,Spinal cord ,Fludarabine ,Translational/Early Phase Clinical Trials ,medicine.anatomical_structure ,Glioma ,Internal medicine ,Toxicity ,Medicine ,AcademicSubjects/MED00300 ,AcademicSubjects/MED00310 ,Neurology (clinical) ,medicine.symptom ,Young adult ,business ,medicine.drug - Abstract
Background We previously discovered high expression of the disialoganglioside GD2 on H3K27M+ gliomas and demonstrated preclinical efficacy of intravenous (IV) GD2-targeted chimeric antigen receptor (CAR) T-cells in preclinical models of H3K27M-mutated diffuse intrinsic pontine glioma (DIPG) and diffuse midline gliomas (DMGs). We are now conducting a Phase I clinical trial (NCT04196413) of autologous GD2-targeting CAR T-cells for H3K27M+ DIPG and spinal cord DMG. Here we present the results of subjects treated at dose level 1 (DL1; 1 million GD2-CAR T-cells/kg IV). Methods Four patients (3 DIPG, 1 spinal DMG; ages 4–25; 1M/3F) were enrolled at DL1. Three subjects with H3K27M+ DIPG received 1e6 GD2-CAR T-cells/kg IV on study. One patient with spinal DMG enrolled but became ineligible after manufacturing and was treated on an eIND at DL1. An Ommaya reservoir was placed in all subjects for therapeutic monitoring of intracranial pressure. Subjects underwent lymphodepletion with fludarabine/cyclophosphamide and remained inpatient for at least two weeks post-infusion. Results All subjects developed cytokine release syndrome (Grade 1–3) manifested by fever, tachycardia and hypotension. Other toxicities included ICANS (Grade 1–2) and neurological symptoms/signs mediated by intratumoral inflammation which we have termed Tumor Inflammation-Associated Neurotoxicity (TIAN). No evidence of on-target, off-tumor toxicity was observed in any patients. No dose-limiting toxicities occurred. CAR T cells trafficked to the CNS and were detected in CSF and blood. 3/4 patients exhibited marked improvement or resolution of neurological deficits and radiographic improvement. The patient treated on an eIND exhibited >90% reduction in spinal DMG volume but progressed by month 3. Re-treatment of this subject via intracerebroventricular administration resulted in a second reduction in spinal DMG volume by ~80%. Conclusions GD2-CAR T-cells at DL1 demonstrate a tolerable safety profile in patients with H3K27M+ DIPG/DMG with clear signs of T-cell expansion and activity including clinical responses.
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- 2021
46. CAR T cells with dual targeting of CD19 and CD22 in adult patients with recurrent or refractory B cell malignancies: a phase 1 trial
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Steven A. Feldman, Kara L. Davis, John H. Baird, Robert Lowsky, Rachel C. Lynn, Magali Bazzano, Judith A. Shizuru, Matthew J. Frank, Surbhi Sidana, Maria Caterina Rotiroti, Maria Iglesias, Sean Mackay, Sally Arai, Bita Sahaf, Shabnum Patel, Nirali N. Shah, Laura Johnston, Jay Y. Spiegel, Jing Zhou, Juliana Craig, Robert S. Negrin, Robbie G. Majzner, Andrew R. Rezvani, Zach Ehlinger, Ilan R. Kirsch, Parveen Shiraz, Chelsea D. Mullins, Michael G. Ozawa, Nikolaos Gkitsas, Crystal L. Mackall, Terry J. Fry, Warren D. Reynolds, Yasodha Natkunam, Sneha Ramakrishna, Scott J. Bornheimer, Allison P. Jacob, Lori Muffly, Jean Oak, Haiying Qin, Katherine A. Kong, Wen-Kai Weng, Everett Meyer, Nasheed Hossain, John S. Tamaresis, Sheren F. Younes, David B. Miklos, Liora M. Schultz, Eric J Yang, and Harshini Chinnasamy
- Subjects
0301 basic medicine ,Oncology ,Adult ,medicine.medical_specialty ,Lymphoma, B-Cell ,Lymphoma ,medicine.medical_treatment ,Sialic Acid Binding Ig-like Lectin 2 ,Antigens, CD19 ,Cancer immunotherapy ,Immunotherapy, Adoptive ,General Biochemistry, Genetics and Molecular Biology ,CD19 ,Article ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,immune system diseases ,Recurrence ,Phase I trials ,Internal medicine ,hemic and lymphatic diseases ,medicine ,Humans ,B-cell lymphoma ,B cell ,Aged ,Acute lymphocytic leukaemia ,biology ,business.industry ,General Medicine ,Immunotherapy ,Middle Aged ,medicine.disease ,Chimeric antigen receptor ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,Disease Progression ,business ,Progressive disease - Abstract
Despite impressive progress, more than 50% of patients treated with CD19-targeting chimeric antigen receptor T cells (CAR19) experience progressive disease. Ten of 16 patients with large B cell lymphoma (LBCL) with progressive disease after CAR19 treatment had absent or low CD19. Lower surface CD19 density pretreatment was associated with progressive disease. To prevent relapse with CD19− or CD19lo disease, we tested a bispecific CAR targeting CD19 and/or CD22 (CD19-22.BB.z-CAR) in a phase I clinical trial (NCT03233854) of adults with relapsed/refractory B cell acute lymphoblastic leukemia (B-ALL) and LBCL. The primary end points were manufacturing feasibility and safety with a secondary efficacy end point. Primary end points were met; 97% of products met protocol-specified dose and no dose-limiting toxicities occurred during dose escalation. In B-ALL (n = 17), 100% of patients responded with 88% minimal residual disease-negative complete remission (CR); in LBCL (n = 21), 62% of patients responded with 29% CR. Relapses were CD19−/lo in 50% (5 out of 10) of patients with B-ALL and 29% (4 out of 14) of patients with LBCL but were not associated with CD22−/lo disease. CD19/22-CAR products demonstrated reduced cytokine production when stimulated with CD22 versus CD19. Our results further implicate antigen loss as a major cause of CAR T cell resistance, highlight the challenge of engineering multi-specific CAR T cells with equivalent potency across targets and identify cytokine production as an important quality indicator for CAR T cell potency., Bispecific CAR T cells targeting CD19 and CD22 exhibit clinical activity and low toxicity in patients with large B cell lymphoma and B cell acute lymphoblastic leukemia, with relapses associated with loss of CD19 but not CD22.
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- 2020
47. Identification of dual positive CD19+/CD3+ T cells in a leukapheresis product undergoing CAR transduction: a case report
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Kara L. Davis, Courtney Erickson, Sneha Ramakrishna, Bita Sahaf, Alice Bertaina, Jean Oak, Robbie G. Majzner, Neehar Bhatia, Steven A. Feldman, Crystal L. Mackall, Christina Baggott, Shabnum Patel, and Liora M. Schultz
- Subjects
Male ,Cancer Research ,pediatrics ,T-Lymphocytes ,medicine.medical_treatment ,Antigens, CD19 ,Immunology ,Population ,receptors ,chemical and pharmacologic phenomena ,Case Report ,Hematopoietic stem cell transplantation ,cell engineering ,adoptive ,CD19 ,immune system diseases ,Acute lymphocytic leukemia ,hemic and lymphatic diseases ,medicine ,Humans ,Immunology and Allergy ,hematologic neoplasms ,Leukapheresis ,Child ,education ,Pharmacology ,education.field_of_study ,Receptors, Chimeric Antigen ,biology ,business.industry ,hemic and immune systems ,Immunotherapy ,Suicide gene ,medicine.disease ,Chimeric antigen receptor ,2518 1619 ,Oncology ,chimeric antigen ,Cancer research ,biology.protein ,Molecular Medicine ,immunotherapy ,business - Abstract
BackgroundChimeric antigen receptor (CAR) therapy and hematopoietic stem cell transplantation (HSCT) are therapeutics for relapsed acute lymphocytic leukemia (ALL) that are increasingly being used in tandem. We identified a non-physiologic CD19+/CD3+ T-cell population in the leukapheresis product of a patient undergoing CAR T-cell manufacturing who previously received a haploidentical HSCT, followed by infusion of a genetically engineered T-cell addback product. We confirm and report the origin of these CD19+/CD3+ T cells that have not previously been described in context of CAR T-cell manufacturing. We additionally interrogate the fate of these CD19-expressing cells as they undergo transduction to express CD19-specific CARs.Main bodyWe describe the case of a preteen male with multiply relapsed B-ALL who was treated with sequential cellular therapies. He received an αβ T-cell depleted haploidentical HSCT followed by addback of donor-derived T cells genetically modified with a suicide gene for iCaspase9 and truncated CD19 for cell tracking (RivoCel). He relapsed 6 months following HSCT and underwent leukapheresis and CAR T-cell manufacturing. During manufacturing, we identified an aberrant T-cell population dually expressing CD19 and CD3. We hypothesized that these cells were RivoCel cells and confirmed using flow cytometry and PCR that the identified cells were in fact RivoCel cells and were eliminated with iCaspase9 activation. We additionally tracked these cells through CD19-specific CAR transduction and notably did not detect T cells dually positive for CD19 and CD19-directed CARs. The most likely rationale for this is in vitro fratricide of the CD19+ ‘artificial’ T-cell population by the CD19-specific CAR+ T cells in culture.ConclusionsWe report the identification of CD19+/CD3+ cells in an apheresis product undergoing CAR transduction derived from a patient previously treated with a haploidentical transplant followed by RivoCel addback. We aim to bring attention to this cell phenotype that may be recognized with greater frequency as CAR therapy and engineered αβhaplo-HSCT are increasingly coupled. We additionally suggest consideration towards using alternative markers to CD19 as a synthetic identifier for post-transplant addback products, as CD19-expression on effector T cells may complicate subsequent treatment using CD19-directed therapy.
- Published
- 2020
48. Locoregionally administered B7-H3-targeted CAR T cells for treatment of atypical teratoid/rhabdoid tumors
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Shaurya Dhingra, Samuel H. Cheshier, Sabine Mueller, Stéphanie Puget, Marcel Kool, Siddhartha Mitra, Derek Yecies, Olivier Delattre, Michael C. Frühwald, Johanna Theruvath, Christopher Mount, Robbie G. Majzner, Sakina Zaidi, Didier Surdez, Louai Labanieh, Daniel Williamson, Poul H. Sorensen, Amaury Leruste, Franck Bourdeaut, Claus Moritz Graef, Alberto Delaidelli, Pascal Johann, Crystal L. Mackall, Michelle Monje, Elena Sotillo, Stefan M. Pfister, Sabine Heitzeneder, Peng Xu, Martin Hasselblatt, and Martina Finetti
- Subjects
Adult ,B7 Antigens ,medicine.medical_treatment ,T-Lymphocytes ,Central nervous system ,Mice, SCID ,medicine.disease_cause ,Cancer Vaccines ,Immunotherapy, Adoptive ,General Biochemistry, Genetics and Molecular Biology ,Article ,Proinflammatory cytokine ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Fetus ,Mice, Inbred NOD ,medicine ,Animals ,Humans ,SMARCB1 ,Receptor ,Cells, Cultured ,Rhabdoid Tumor ,030304 developmental biology ,Injections, Intraventricular ,0303 health sciences ,Chemotherapy ,Receptors, Chimeric Antigen ,business.industry ,Brain Neoplasms ,Teratoma ,Brain ,Infant ,General Medicine ,Immunotherapy ,Xenograft Model Antitumor Assays ,3. Good health ,Radiation therapy ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Child, Preschool ,Cancer research ,Female ,business ,Carcinogenesis - Abstract
Atypical teratoid/rhabdoid tumors (ATRTs) typically arise in the central nervous system (CNS) of children under 3 years of age. Despite intensive multimodal therapy (surgery, chemotherapy and, if age permits, radiotherapy), median survival is 17 months(1,2). We show that ATRTs robustly express B7-H3/CD276 that does not result from the inactivating mutations in SMARCB1 (refs.(3,4)), which drive oncogenesis in ATRT, but requires residual SWItch/Sucrose Non-Fermentable (SWI/SNF) activity mediated by BRG1/SMARCA4. Consistent with the embryonic origin of ATRT(5,6), B7-H3 is highly expressed on the prenatal, but not postnatal, brain. B7-H3. BB.z-chimeric antigen receptor (CAR) T cells administered intracerebroventricularly or intratumorally mediate potent antitumor effects against cerebral ATRT xenografts in mice, with faster kinetics, greater potency and reduced systemic levels of inflammatory cytokines compared to CAR T cells administered intravenously. CAR T cells administered ICV also traffic from the CNS into the periphery; following clearance of ATRT xenografts, B7-H3.BB.z-CAR T cells administered intracerebroventricularly or intravenously mediate antigen-specific protection from tumor rechallenge, both in the brain and periphery. These results identify B7-H3 as a compelling therapeutic target for this largely incurable pediatric tumor and demonstrate important advantages of locoregional compared to systemic delivery of CAR T cells for the treatment of CNS malignancies.
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- 2020
49. Tuning the Antigen Density Requirement for CAR T Cell Activity
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Aidan Tousley, Sabine Heitzeneder, Johanna Theruvath, Robbie G. Majzner, Skyler P. Rietberg, Elena Sotillo, Volker Wiebking, Ronald D. Vale, Rui Dong, Rebecca Richards, Peng Xu, Evan W. Weber, Alexander R. Dunn, Rachel C. Lynn, Louai Labanieh, Vipul T. Vachharajani, Sang M. Nguyen, Meena Kadapakkam, Crystal L. Mackall, and June Helen Myklebust
- Subjects
0301 basic medicine ,Cell signaling ,Oncology and Carcinogenesis ,CD19 ,Article ,Synapse ,Vaccine Related ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Antigen ,Receptors ,medicine ,Animals ,Humans ,Tyrosine ,Cancer ,Receptors, Chimeric Antigen ,biology ,Chemistry ,Chimeric Antigen ,medicine.disease ,Chimeric antigen receptor ,Lymphoma ,Cell biology ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,biology.protein ,Immunization ,human activities ,CD8 ,Signal Transduction - Abstract
Insufficient reactivity against cells with low antigen density has emerged as an important cause of chimeric antigen receptor (CAR) T-cell resistance. Little is known about factors that modulate the threshold for antigen recognition. We demonstrate that CD19 CAR activity is dependent upon antigen density and that the CAR construct in axicabtagene ciloleucel (CD19-CD28ζ) outperforms that in tisagenlecleucel (CD19-4-1BBζ) against antigen-low tumors. Enhancing signal strength by including additional immunoreceptor tyrosine-based activation motifs (ITAM) in the CAR enables recognition of low-antigen-density cells, whereas ITAM deletions blunt signal and increase the antigen density threshold. Furthermore, replacement of the CD8 hinge-transmembrane (H/T) region of a 4-1BBζ CAR with a CD28-H/T lowers the threshold for CAR reactivity despite identical signaling molecules. CARs incorporating a CD28-H/T demonstrate a more stable and efficient immunologic synapse. Precise design of CARs can tune the threshold for antigen recognition and endow 4-1BBζ-CARs with enhanced capacity to recognize antigen-low targets while retaining a superior capacity for persistence. Significance: Optimal CAR T-cell activity is dependent on antigen density, which is variable in many cancers, including lymphoma and solid tumors. CD28ζ-CARs outperform 4-1BBζ-CARs when antigen density is low. However, 4-1BBζ-CARs can be reengineered to enhance activity against low-antigen-density tumors while maintaining their unique capacity for persistence. This article is highlighted in the In This Issue feature, p. 627
- Published
- 2020
50. Transient 'rest' induces functional reinvigoration and epigenetic remodeling in exhausted CAR-T cells
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Howard Y. Chang, Peng Xu, Kevin R. Parker, Thomas J. Wandless, Zinaida Good, Elena Sotillo, Hima Anbunathan, Yanyan Qi, Robbie G. Majzner, Panayiotis Vandris, Ansuman T. Satpathy, Ling-chun Chen, Rachel C. Lynn, Crystal L. Mackall, Evan W. Weber, Andrew J. Gentles, Meena Malipatlolla, and John Lattin
- Subjects
0303 health sciences ,Cell signaling ,T cell ,Cell ,Biology ,complex mixtures ,Cell biology ,Dasatinib ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,medicine ,Epigenetics ,human activities ,Transcription factor ,Reprogramming ,030304 developmental biology ,medicine.drug ,Proto-oncogene tyrosine-protein kinase Src - Abstract
SUMMARYT cell exhaustion limits immune responses against cancer and is a major cause of resistance to CAR-T cell therapeutics. Using a model wherein tonic CAR signaling induces hallmark features of exhaustion, we employed a drug-regulatable CAR to test the impact of transient cessation of receptor signaling (i.e. “rest”) on the development and maintenance of exhaustion. Induction of rest in exhausting or already-exhausted CAR-T cells resulted in acquisition of a memory-like phenotype, improved anti-tumor functionality, and wholescale transcriptional and epigenetic reprogramming. Similar results were achieved with the Src kinase inhibitor dasatinib, which reversibly suppresses CAR signaling. The degree of functional reinvigoration was proportional to the duration of rest and was associated with expression of transcription factors TCF1 and LEF1. This work demonstrates that transient cessation of CAR-T cell signaling can enhance anti-tumor potency by preventing or reversing exhaustion and challenges the paradigm that exhaustion is an epigenetically fixed state.
- Published
- 2020
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